Cloning of the Protective Antigen Gene of Bacillus anthracis
Abstract
The tripartite protein toxin of Bacillus anthracis consists of protective antigen (PA), edema factor (EF), and lethal factor (LF). As a first step in developing a more efficacious anthrax vaccine, recombinant plasmids containing the PA gene have been isolated. A library was constructed in the E. coli vector pBR322 from Bam HI-generated fragments of the anthrax plasmid, pBA1. Two clones producing PA were identified by screening lysates with ELISA (enzyme-linked immunosorbent assay). Westem blots revealed a full-size PA protein in the recombinant E. coli, and a cell elongation assay demonstrated biological activity. Both positive clones had a 6 kb insert of DNA, which mapped in the Bam Hi site of the vector. The two inserts are the same except that they lie in opposite orientations with respect to the vector. Thus PA is encoded by the plasmid pBA1.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 1983
- Accession Number
- ADA133885
Entities
People
- Michael H. Vodkin
- Stephen H. Leppla
Organizations
- United States Army Medical Research Institute of Infectious Diseases